SW 1353 cells were grown in 96-well plates and then incubated for 24 h in serum-free medium supplemented with 500 μM DMOG, 1 µM doxorubicin or DMSO as a negative control. Then, cell viability was assessed using a Cell Titer Glo 2.0 kit (Promega, Fitchburg, WI, USA), which determines the number of viable cells in a culture by quantifying intracellular ATP, according to the manufacturer’s instruction. Finally, cells were grown until 90% confluence in 6-well plates and were then treated with 500 μM DMOG, 1 µM doxorubicin or DMSO for 24 h in serum-free DMEM. Secretion of GAPDH in these cells was evaluated by western blotting as previously described.
Celltiter glo 2.0 kit
Manufactured by Promega
Sourced in United States
The CellTiter-Glo 2.0 kit is a cell viability assay that measures the presence of ATP, an indicator of metabolically active cells. The kit uses a luciferase-based detection system to quantify the ATP levels in a sample, providing a luminescent readout that is proportional to the number of viable cells present.
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Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Synergy neo, by Agilent Technologies (2 mentions)
Celltiter glo 2, by Promega (2 mentions)
Adenosine triphosphate (atp), by Merck Group (2 mentions)
Optima plate reader, by BMG Labtech (2 mentions)
H4 plate reader, by Agilent Technologies (2 mentions)
Enspire 2300, by Promega (2 mentions)
Cytotox 96 non radioactive cytotoxicity assay kit, by Promega (2 mentions)
Spark plate reader, by Tecan (2 mentions)
Victor multilabel plate reader, by PerkinElmer (1 mentions)
Spark multimode microplate reader, by Tecan (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Carl Roth (1 mentions)
Celltiter glo 3d kit, by Promega (1 mentions)
96 well plate, by Corning (1 mentions)
Centro lb 960, by Berthold Technologies (1 mentions)
Nad nadh glo assay, by Promega (1 mentions)
Crystal violet, by Thermo Fisher Scientific (1 mentions)
Biotek synergy h1 microplate reader, by Agilent Technologies (1 mentions)
Pherastar, by BMG Labtech (1 mentions)
43 protocols using celltiter glo 2.0 kit
1
DMOG and Doxorubicin Effects on Cell Viability and GAPDH Secretion
2
Extracellular ATP as Cell Damage Indicator
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The integrity of the cell membrane can be damaged by antibiotics or YH68, accompanied by leakage of intracellular ATP. Therefore, changes in extracellular ATP levels can accurately reflect cell damage. The extracellular ATP level was measured at appropriate time intervals by a previously described bioluminescence-based method (Suzuki et al., 2005 (link)) using a microplate reader (PE EnSpire 2300) and a luminescence kit (CellTiter-Glo® 2.0 Kit, Promega, United States).
3
Cytotoxicity Evaluation of BBR and OLX
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Vero E6 cells that were seeded in 96-well plates at a density of 3 × 104 cells/well were treated with two-fold dilution series of BBR and OLX in the absence of SARS-CoV-2 infection. Cells treated with 0.1% DMSO were used as a negative control. At 24 h post-treatment, the cell viability was assessed using the Cell Titer Glo 2.0 kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions. Luminescence was detected on the Victor Multilabel Plate Reader (Perkin Elmer, Waltham, MA, USA). The 50% cytotoxicity concentration (CC50) value was estimated by four parameter logistic regression of the data, using Graphpad Prism (version 5.0).
4
ALS Motor Neuron Survival Assay
5
Cytotoxicity and Cell-Cell Fusion Assay
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The cytotoxicity assay was performed using the CellTiter-Glo 2.0 kit (Promega) to measure the cell viability (changes in the amount of ATP due to cell death) when exposed to different compounds. Another identical set of Env-expressing cells, CD4- and CCR5-expressing cells and compounds were mixed in parallel when the cell-cell fusion assay was performed, followed by incubation at 37°C for 2 hr. The cells were cooled to room temperature for 30 min before adding 100 μl of the CellTiter-Glo 2.0 reagent. The mixture was incubated in room temperature in dark for 10 min before recording luminescence using a Synergy Neo microplane reader (BioTek).
6
Cytotoxicity and Cell-Cell Fusion Assay
7
Intracellular and Extracellular ATP Measurement
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To measure the ATP concentration, we used the CellTiter-Glo 2.0 kit (Promega) and followed the manufacturer’s instruction. In brief, S2 cells (8×104) were plated in a transparent 96-well plate and treated as indicated above. Every treatment was performed at least in triplicate and each experiment was performed two or three times. To determine the intracellular ATP concentration, 100 μl of fresh incubation medium was added to the treated cells and a 0.5× volume CellTiter-Glo 2.0 (Promega) was added to each well. The background signal was determined by adding CellTiter-Glo 2.0 (Promega) to medium without cells. The total volume was transferred to a 96-well Greiner flat white plate. Luminescence was measured with a Tecan Spark multimode microplate reader (emission wavelength 560 nm).
To determine the extracellular ATP level, the incubation medium was transferred to a 96-well Greiner flat white plate and 0.5× volume CellTiter-Glo 2.0 (Promega) was added to each well. The luminescence was measured as above. An ATP calibration curve was made by measuring different known concentrations of ATP (Sigma-Aldrich) in Schneider's medium.
To determine the extracellular ATP level, the incubation medium was transferred to a 96-well Greiner flat white plate and 0.5× volume CellTiter-Glo 2.0 (Promega) was added to each well. The luminescence was measured as above. An ATP calibration curve was made by measuring different known concentrations of ATP (Sigma-Aldrich) in Schneider's medium.
8
Cytotoxicity Assessment of MTT Assay
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MTT (Sigma-Aldrich) solution was added to the RAW 264.7 cell culture to a final concentration of 500 μg/ml and incubated for 1–2 h (37 °C, 5% CO2). After discarding the medium, MTT was solubilized in 100 µl of dimethyl sulfoxide (DMSO) (Roth) per well (15 min, room temperature) and the absorbance was measured on a microplate absorbance reader (Sunrise Tecan Reader) at 562 nm. The decrease in cell respiration (%) was calculated as 100 × (1 − (OD exp. mean value (− substrate blank)/OD control mean value (− substrate blank)). Cellular ATP measurements were performed using the CellTiter-Glo 2.0 Kit (Promega) according to the manufacturer’s instructions.
9
Keratinocyte Viability Assay
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Skin-derived keratinocytes were trypsinized, counted on a hemocytometer, and seeded on a white background plate at a density of 5,000 per well in triplicate. These cells grew for 7 days at 37° C in a humidified chamber with 5% carbon dioxide. After 0,2,4, and 6 days, a luminescent cell viability assay was conducted using CellTiter-Glo 2.0 kit (Promega, Cat# G9242) according to the manufacturer’s instructions. Fluorescence intensity was measured at 485–500nmEx/520–530nmEm.
10
Cell Viability Assay for Neuroblastoma
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Cells were plated on 96-well plates at a density of 10,000 (SK-N-BE(2)-C, IMR-32, Kelly) or 20,000 (NB8) cells per well and treated as indicated for 72 h. According to manufacturer’s protocol, cells were incubated with reagent for 25 min using the CellTiter-Glo 2.0 kit (Promega, SK-N-BE(2)-C, IMR-32, Kelly) or the CellTiter-Glo 3D kit (Promega, NB8) and bioluminescence was read in an OPTIMA plate reader (BMG Labtech).
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